CN111690763A - DNA bar code for identifying variety of Chinese wolfberry and identification method thereof - Google Patents
DNA bar code for identifying variety of Chinese wolfberry and identification method thereof Download PDFInfo
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Abstract
The invention discloses a DNA bar code for identifying lycium species and an identification method thereof, belonging to the technical field of lycium species identification. The invention can identify the lycium barbarum, the lycium ruthenicum murr, the lycium barbarum with the germplasm name of 9001 lycium barbarum, the lycium barbarum of Ningxia Huangguosha, the lycium changii, the lycium ruthenicum murr and the like by utilizing the DNA bar code of trnG-trnS. The DNA bar code provided by the invention can be applied to the construction of a medlar phylogenetic tree, and further can be used for researching the phylogeny of medlar intra-species and inter-species, and provides an effective basis for variety identification, classification and phylogeny research of medlar. The trnG-trnS barcode database is obtained on the basis of the identification method, the trnG-trnS sequence of the sample to be identified is compared with the sequence in the trnG-trnS barcode database, the species of the medlar can be effectively identified, the interspecific relationship of the medlar is determined, and effective basis is provided for the variety of the medlar.
Description
Technical Field
The invention belongs to the technical field of lycium species identification, and particularly relates to a DNA bar code for lycium species identification and an identification method thereof.
Background
The medlar is a plant of the Lycium of Solanaceae (Solanaceae) Solanum (Solanae Reichb.) Lycium subfamily (Lycineae Wettst), is perennial deciduous shrub, and has high medicinal and health care values of fruits, root barks and leaves. About 80 types of the plants belong to a world distribution, and the domestic main planting areas are distributed in Ningxia, Xinjiang, Gansu, Qinghai and other places. Because the varieties have the same primitive or similar reason and have high similarity of development morphology, tissue structure and chemical components, the traditional morphological identification method has difficult to solve the problem.
The DNA barcode (DNA barcoding) is a new technology for identifying and identifying target species by using 1 or a few DNA fragments, has the characteristics of simple and convenient operation, high accuracy, quick identification and the like, and has become an interesting new direction and research hotspot in modern biological taxonomy research. In recent years, researchers at home and abroad have conducted active research and study on DNA barcode gene sequences suitable for identifying plants.
The invention patent with publication number CN110229927A, which is a method for identifying lycium ruthenicum based on DNA bar codes and application thereof, provides a method for identifying lycium ruthenicum based on DNA bar codes. The DNA barcode gene sequence for identifying Lycium ruthenicum Murr is LRITS2(the inside transcribed spacer of the second ribosomal RNA)/LRpsbA-trnH (a non-coding region between the chloroplast genes psbA and trnH). The DNA barcode sequences LRITS2/LRpsbA-trnH for identifying Lycium ruthenicum can be used simultaneously or optionally. The method can efficiently and accurately identify the lycium ruthenicum raw material. The method can prevent similar confused products or fake products, can be used for identifying fruit powder, fruit powder and the like, and has important application value and great social benefit for guaranteeing food safety and consumer rights.
An article "early screening of matrimony vine intraspecific hybrids based on ITS barcode sequences" discloses the early screening study of matrimony vine hybrids using the ribose Internal Transcribed Spacer (ITS) barcode sequences. Extracting DNA of the leaves of the Chinese wolfberry by adopting an improved Cetyl Trimethyl Ammonium Bromide (CTAB) method, amplifying and cloning nrDNAITS (negative deoxyribose nucleic acid) regions in the DNA by utilizing a synthesized specific primer, and sequencing and analyzing a target fragment. The results show that the variety Ningqi No.1, Ningqi No. 2 and white flower wolfberry in Ningxia wolfberry species are used as male parent and female parent, hybridization combination is selected, clustering analysis is carried out on hybrid progeny generated by intraspecific hybridization breeding based on ITS barcode sequences, the genetic relationship and difference between the hybrid progeny and the male parent and the female parent are analyzed, and early screening is carried out on the hybrid progeny. The result shows that the ITS-based barcode sequence can be used as one of the methods for early screening hybrid breeding offspring, and has important significance for establishing a molecular marker-assisted breeding technology and shortening the breeding cycle. However, the existing barcode database of lycium is relatively deficient in species abundance and quantity, and still needs a lot of research.
Disclosure of Invention
The invention provides a rapid identification method of molecular markers of representative lycium barbarum with germplasm in local varieties of Ningxia lycium barbarum, northern, Xinjiang, Yunnan, Hebei and the like, hybrid populations, space mutation populations, ploidy populations and other different lycium barbarum sources, and can be applied to identification of lycium barbarum species by relying on unique lycium barbarum germplasm resource gardens in China and breeding materials obtained by carrying out breeding of new lycium barbarum varieties for a long time.
The invention aims to provide a method for identifying variety and determining interspecies relationship of medlar by using DNA (deoxyribonucleic acid) bar codes, and meanwhile, a trnG-trnS bar code database is obtained on the basis of the method, and by comparing trnG-trnS sequences of a sample to be identified with sequences in the trnG-trnS bar code database, the variety and the interspecies relationship of medlar can be effectively identified, so that an effective basis is provided for variety of medlar.
The technical scheme of the invention is as follows:
the first purpose of the invention is to provide a method for identifying the variety of the medlar by using a DNA bar code, wherein the DNA bar code is a trnG-trnS bar code.
The identifiable species of Lycium barbarum include Lycium barbarum No.1 (l.barbarbarbarum lin), Lycium barbarum No. 2, Lycium barbarum No.3, Lycium barbarum No. 4, Lycium barbarum No. 5, Lycium barbarum No. 6, Lycium ningbarl No. 7, Lycium ningnong No. 9, Lycium xanthophyllum barbarum (l.barbarbarum lin. var. aurantianum k. f. china var. nov.nov.), Lycium barbarum (Lycium barbarum), Lycium nigrum ruthenicum (Lycium ruthenicum Murr.), Lycium barbarum No. 5 (l.barbarbarbarbarbarbarum lin), Lycium barbarum (Lycium barbarum l. var. minium) poinanum (Pojark. a. m. lu.), Lycium barbarum (lybarum barbarum), Lycium barbarum (l.barbararum. barbarum), Lycium barbarum (l.1), Lycium barbarum (Lycium barbarum), Lycium barbarum (l.1. yunnary mutation), Lycium barbarum (l. yunnary mutation), Lycium barbarum (l.1. yunnary stem) Round fruit medlar, variety name is 9001 medlar, Ningxia Huangguo medlar, Changji medlar, Hebei medlar, etc.
Preferably, the method for identifying the variety of the Chinese wolfberry based on the DNA bar code comprises the following steps:
1) extracting genome DNA from a medlar sample;
2) amplifying a sequence fragment of the trnG-trnS bar code by using the genomic DNA as a template and primers shown as nucleotide sequences SEQ ID NO.37 and SEQ ID NO.38 to obtain a PCR product;
3) sequencing the PCR amplification product;
4) and (5) constructing a phylogenetic tree and identifying the medlar.
The invention carries out contraposition arrangement through screening the obtained trnG-trnS sequences, carries out analysis and calculation on the base composition of a target sequence, the base variation frequency among the sequences, the conversion and inversion frequency among the sequences and the ratio thereof, compares the distribution of the intra-species and inter-species differences of the sequences, constructs a phylogenetic tree, and establishes an identification technical system of the trnG-trnS DNA bar code for identifying the Chinese wolfberry varieties.
Preferably, in the step 1), the DNA extracted from the sample is extracted by using a kit.
More preferably, in the step 1), the Kit is a DNA secure Plant Kit.
More preferably, the step of DNA extraction using the kit is as follows:
(1) extraction of DNA
Collecting fresh and tender leaves of fructus Lycii plant as sample, washing, and storing at-80 deg.C. The total DNA is extracted by adopting a novel Plant genome DNA extraction Kit (DNA sure Plant Kit), and the extraction method comprises the following steps:
firstly, 100g of sample is ground in a multifunctional high-efficiency biological sample preparation instrument for 22 times/s for 2 min. 400ul of buffer LP1 and 6ul of RNase A (10mg/ml) were immediately added thereto, vortexed for 1min, and allowed to stand at room temperature for 10 min.
② adding 130ul buffer solution LP2, mixing well, and vortex shaking for 1 min.
③ centrifuging at 12000rpm for 5min, and transferring the supernatant to a new centrifuge tube.
Adding 1.5 times of buffer LP3 (testing whether absolute ethyl alcohol is added before use), immediately and fully shaking and mixing for 15sec, wherein flocculent precipitates may appear.
Fifthly, adding the solution and flocculent precipitate obtained in the previous step into an adsorption column CB3 (placing the adsorption column into a collecting pipe), centrifuging at 12000rpm for 30s, pouring the waste liquid, and placing an adsorption column CB3 into the collecting pipe.
Sixthly, 600ul of rinsing liquid PW (whether absolute ethyl alcohol is added in the adsorption column CB3 is checked before use) is added into the adsorption column CB3, the adsorption column CB3 is placed into a collecting pipe after the adsorption column CB3 is centrifuged at 12000rpm for 30 s. (Note: if the adsorption column membrane is green, 500ul of absolute ethanol was added to adsorption column CB3, centrifuged at 12000rpm for 30s, the waste liquid was discarded, and adsorption column CB3 was put into the collection tube)
And seventhly, repeating the operation step 6.
And eighthly, putting the adsorption column CB3 back into the collecting pipe, centrifuging at 12000rpm for 2min, and pouring the waste liquid. The adsorption column CB3 was left at room temperature for 15min to thoroughly dry the residual rinse solution in the adsorption material.
Ninthly, transferring the adsorption column CB3 into a clean centrifugal tube, hanging and dripping 100ul of elution buffer TE into the middle part of the adsorption film, placing the adsorption film at room temperature for 2min, centrifuging the adsorption film at 12000rpm for 2min, and collecting the solution into the centrifugal tube.
(2) DNA concentration and purity measurement
Firstly, agarose gel electrophoresis detection
1.2% agarose gel was prepared with 1.2g agarose and 100ml 1 × TAE buffer, and 4ul ddH was added to the PCR tube2The detection system of O +1ul DNA sample (undiluted) +1ul 6 loading buffer was subjected to agarose gel electrophoresis. And observing the detection result under an ultraviolet gel imaging system.
② ultraviolet spectrophotometer detection
The ultraviolet spectrophotometer is preheated in advance,add 99ul ddH to PCR tube2O +1ul DNA samples (undiluted) were tested. The detection result shows the concentration and OD of the sample260/OD280Ratio, OD260/OD280The ratio should be 1.7-1.9, if elution is performed without using elution buffer, ddH is used2O, ratio is low because the pH and the presence of ions affect the light absorption but do not indicate low purity.
Preferably, in the step 2), the reaction system for PCR amplification is: firstly, pre-denaturation is carried out for 2min at 94 ℃; ② denaturation at 94 ℃ for 30s, annealing at 55 ℃ for 30s (the annealing temperature can be adjusted between 58 ℃ and 60 ℃), extension at 72 ℃ for 2min, and 35 cycles; ③ keeping the temperature at 72 ℃ for 10 min; and fourthly, storing at 4 ℃. And (3) detecting the PCR product by 1.0% agarose gel electrophoresis, and observing the amplification result under an ultraviolet gel imaging system.
Preferably, in the step 3), the sequencing step is as follows:
(1) cloning of PCR products:
and (3) recovering the target band by using an AxyPrep DNA gel recovery kit, recovering and detecting by using 1.2% agarose gel electrophoresis, and taking the purified target DNA as a sequencing template. The recovered product was ligated to a T vector (pGEM-T) using pLB zero background rapid cloning kit, and then transformed into E.coli DH 5. alpha. for culture. And screening positive colonies by adopting a blue-white spot method, and carrying out colony PCR detection. The amplification results were observed under a uv gel imaging system.
(2) Sequencing and analysis of the sequence:
the colonies of positive clones were subjected to DNA sequencing and to homology alignment with the sequences published in NCBI and the sequences were analyzed. The specific operation is as follows:
after PCR detection is carried out on positive colonies, the colonies containing target fragments are cultured by an LB liquid culture medium, 3 colonies are selected from each material and sent to a company Limited in Biotechnology engineering (Shanghai) to be sequenced by a Sanger method, and a trnG-trnS sequence is obtained.
And carrying out homology comparison on the obtained DNA barcode gene sequence and published sequences in an NCBI database by using DNAMAN, and respectively carrying out alignment arrangement on the DNA barcode gene sequence of the medlar by using a Clustal X program. The base composition of the target sequence, the frequency of base variation between sequences and the frequency and ratio of transition transversions between sequences were calculated using the phylogenetic analysis software MEGA 7.0. By comparing the distribution of sequence intraspecific and interspecific differences, phylogenetic trees are constructed. And establishing a trnG-trnS bar code database for the identification of the Chinese wolfberry variety.
The second purpose of the invention is to provide a trnG-trnS barcode database of the medlar sample, which is constructed by the method, and comprises 36 trnG-trnS barcodes, wherein the nucleotide sequence of the trnG-trnS barcode is shown in SEQ ID NO. 1-36.
The third purpose of the invention is to provide the application of identifying the variety of the medlar based on the trnG-trnS bar code database of the medlar sample.
Preferably, the application of identifying the variety of the Chinese wolfberry based on the trnG-trnS bar code database of the Chinese wolfberry sample comprises the following steps:
and comparing the trnG-trnS bar code sequence of the sample to be identified with the sequences in the trnG-trnS bar code database to identify the Chinese wolfberry variety.
By comparing the trnG-trnS sequence of the sample to be identified with the sequence in the trnG-trnS bar code database of the medlar sample, the species of medlar can be effectively identified, the interspecific relationship of medlar can be determined, and an effective basis is provided for classification and identification of the variety of medlar by determining the interspecific relationship between the medlar to be detected and the medlar in the bar code database.
Preferably, the trnG-trnS barcode sequence of the sample to be identified is obtained by extracting genomic DNA, performing PCR amplification and sequencing PCR amplification products to obtain a corresponding sequence, and the operation steps are the same as the steps 1), 2) and 3) in the method for identifying the variety of the Chinese wolfberry by using the DNA barcode.
The invention has the beneficial effects that:
(1) the identification method of the Chinese wolfberry variety is established by utilizing the trnG-trnS bar code gene for the first time. Can be used for identifying the varieties of Ningqi, Huangguoyi, Heiguo, northern, hemp leaf, Chinese, Yunnan, vine, purple-stalk and red-branch Lycium.
(2) The DNA bar code provided by the invention can be applied to the construction of a medlar phylogenetic tree, and further used for researching the phylogeny of medlar intra-species and inter-species, and the genetic diversity and genetic relationship of medlar are disclosed based on trnG-trnS gene. Meanwhile, an effective basis is provided for variety identification, classification and phylogenetic research of the medlar.
(3) The variety of the Chinese wolfberry is accurately identified based on the trnG-trnS gene.
(4) The trnG-trnS bar code database of the medlar sample is established based on the identification method, and comprises local varieties of Ningxia medlar, local varieties of North, Xinjiang, Yunnan, Hebei, Sichuan and the like, and simultaneously comprises varieties of Lycium ruthenicum Murr, Huangguo medlar, round fruit medlar, red branch medlar and the like, and different medlar sources of hybrid groups, space mutation groups, ploidy groups and the like, and are the medlar with representative germplasm in the whole country, so that an effective basis can be provided for classification and identification of medlar varieties.
By comparing the trnG-trnS bar code of the sample to be identified with the sequence in the trnG-trnS bar code database of the medlar sample, the medlar species can be effectively identified, the interspecific relationship of medlar is determined, and an effective basis is provided for classification and identification of medlar species.
Drawings
Fig. 1 is a DNA extraction detection result of a lycium barbarum sample according to embodiment 1 of the present invention, where a lane M: marker (DL2000DNA molecular marker), wherein a figure (a) shows the DNA extraction detection result of the Chinese wolfberry variety numbers 1-24 in the embodiment 1 of the invention; FIG. b shows the results of DNA extraction and detection of the variety Nos. 25 to 36 of Lycium barbarum of example 1 of the present invention.
FIG. 2 shows the PCR amplification result of trnG-trnS sequence of a portion of the Lycium barbarum sample of example 1 of the present invention, wherein lane M: marker (DL2000DNA molecular marker), lanes 1-4: all the PCR products of the hemp leaves of the invention are the 26 No. of the embodiment 1; 1-4 PCR validation was repeated four times for the samples.
FIG. 3: example 1 of the present invention is a test chart of the trnG-trnS sequence cloning results for Lycium barbarum sample No. 26 cannabis leaves, wherein M: marker (DL2000DNA molecular marker), lane 1: positive cloning; lane 2: negative cloning; lanes 3-9: positive clones (multiple replicates).
FIG. 4: the NJ phylogenetic tree is constructed by trnG-trnS barcodes in a trnG-trnS barcode database of a medlar sample in the embodiment 1 of the invention.
FIG. 5: the invention provides an NJ phylogenetic tree of a part of trnG-trnS bar codes in a wolfberry sample and a database to be identified in experimental example 1.
FIG. 6: the invention is an NJ phylogenetic tree of part of trnG-trnS barcodes in a wolfberry sample and a database to be identified in experimental example 2.
Detailed Description
The invention is described below by means of specific embodiments. Unless otherwise specified, the technical means used in the present invention are well known to those skilled in the art. In addition, the embodiments should be considered illustrative, and not restrictive, of the scope of the invention, which is defined solely by the claims. It will be apparent to those skilled in the art that various changes or modifications in the components and amounts of the materials used in these embodiments can be made without departing from the spirit and scope of the invention. The present invention is further illustrated by the following specific examples.
Example 1 identification of Lycium barbarum samples and construction of a trnG-trnS Bar code database of Lycium barbarum samples
The invention will be further illustrated by the following examples.
1. Samples of trnG-trnS barcode database of wolfberry samples
In order to construct the trnG-trnS barcode database of the wolfberry samples, 36 wolfberry samples from different regions and with similar shapes are collected, and the details are shown in Table 1:
TABLE 1 Lycium plant samples (Lycium samples trnG-trnS Bar code database)
(Note: variety No. 15-20, 22, 33, 35 from open literature 1: "all open varieties in Wan, Yajun, An Wei, et al. identification of 21 parts of Lycium plants [ J ]. Jiangsu Agricultural Science, 2019, 47(01): 64-67.", variety No. 13 from open literature 2: "Shiquan, An Wei, Pyrenin, et al. a new method for identifying Lycium barbarum for vegetable use-nrDNA ITS sequencing (English) [ J ]. Agricutural Science & Technology (2):64-65+ 111." Table 1; variety No. 24 from open literature 3: "Shigang Shiquan., 18 parts of Ningxia Lycium barbarum resources based on nrDNA sequences [ J ]. Anhui Agricultural Science (24, 10379-: 10379-; variety number 10 is derived from document 5: "Yijun, An Wei, Zhao Jianhua, etc.. Lycium ruthenicum transcriptome SSR information analysis and molecular marker development [ J ]. proceedings of Zhejiang university of agriculture and forestry, 2019,36(02):215 + 221."; number 25 resource number: 1111C0003121000044, classification code: 11132115107 from Baidu encyclopedia)
2. Identification method of wolfberry sample and construction method of trnG-trnS bar code database of wolfberry sample
1) Extraction of DNA
Collecting the fresh and tender leaves of the 36 parts of Chinese wolfberry plants in a 5ml freezing tube at the base of the research institute of Chinese wolfberry engineering technology of Ningxia agriculture and forestry academy of sciences, immediately placing the leaves into liquid nitrogen after marking, and preserving the leaves at minus 80 ℃. Sampling time: 6 months 2018, sampling site: the germplasm resource library of national forest trees of medlar in Ningxia Yichuan city is shown in table 1 in detail.
The total DNA is extracted by adopting a novel Plant genome DNA extraction Kit (DNA sure Plant Kit), and the extraction method comprises the following steps:
(1) 100g of the sample was ground in a multifunctional high performance biological sample preparation apparatus for 2min (22 query/s;). 400ul of buffer LP1 and 6ul of RNase A (10mg/ml) were immediately added thereto, vortexed for 1min, and allowed to stand at room temperature for 10 min.
(2) Adding 130ul buffer LP2, mixing well, and vortex shaking for 1 min.
(3) Centrifuge at 12000rpm for 5min and transfer the supernatant to a new centrifuge tube.
(4) After 1.5 times the volume of the buffer LP3 was added (before use, whether or not absolute ethanol was added was checked), the mixture was immediately and thoroughly shaken and mixed for 15 seconds, and at this time, a flocculent precipitate may appear.
(5) Adding the solution and flocculent precipitate obtained in the previous step into an adsorption column CB3 (the adsorption column is placed into a collecting pipe), centrifuging at 12000rpm for 30s, pouring off waste liquid, and placing an adsorption column CB3 into the collecting pipe.
(6) 600ul of the rinsing liquid PW (before use, whether or not absolute ethyl alcohol was added was checked) was added to the adsorption column CB3, the mixture was centrifuged at 12,000rpm for 30 seconds, the waste liquid was discarded, and the adsorption column CB3 was put into a collection tube. (Note: if the adsorption column membrane is green, 500ul of absolute ethanol was added to adsorption column CB3, centrifuged at 12,000rpm for 30s, the waste liquid was discarded, and adsorption column CB3 was put into the collection tube)
(7) Operation 6 is repeated.
(8) The adsorption column CB3 was returned to the collection tube, centrifuged at 12,000rpm for 2min, and the waste liquid was discarded. The adsorption column CB3 was left at room temperature for 15min to thoroughly dry the residual rinse solution in the adsorption material.
(9) Transferring the adsorption column CB3 into a clean centrifuge tube, suspending 100ul of elution buffer TE dropwise into the middle part of the adsorption membrane, standing at room temperature for 2min, centrifuging at 12,000rpm for 2min, and collecting the solution into the centrifuge tube.
(10) Operation 9 is repeated. The DNA product was stored at-80 ℃. To prevent DNA degradation.
2) DNA concentration and purity measurement
Firstly, agarose gel electrophoresis detection
1.2% agarose gel was prepared with 1.2g agarose and 100ml 1 × TAE buffer, and 4ul ddH was added to the PCR tube2The detection system of O +1ul DNA sample (undiluted) +1ul 6 loading buffer was subjected to agarose gel electrophoresis. The results were observed under a UV gel imaging system, as shown in FIG. 1.
② ultraviolet spectrophotometer detection
Preheating ultraviolet spectrophotometer in advance, adding into PCR tube99ul ddH2O +1ul DNA samples (undiluted) were tested. The detection result shows the concentration and OD of the sample260/OD280Ratio, OD260/OD280The ratio should be 1.7-1.9, if elution is performed without using elution buffer, ddH is used2O, ratio is low because the pH and the presence of ions affect the light absorption but do not indicate low purity.
3) PCR amplification
Adding reagents required by amplification such as primers and the like into the DNA obtained in the step 1) as a template to perform PCR amplification. Specific primers and amplification systems are shown in tables 2 and 3.
(1) Primers were designed as follows:
TABLE 2 Universal primers for DNA barcode gene trnG-trnS
(2) PCR amplification System:
the genomic DNA of the test material was subjected to PCR amplification using the above primers, and the amplification system is shown in Table 3:
TABLE 3 DNA Bar code reaction System
The PCR reaction program is that firstly, the mixture is pre-denatured for 2min at 94 ℃; ② denaturation at 94 ℃ for 30s, annealing at 55 ℃ for 30s (the annealing temperature can be adjusted between 58 ℃ and 60 ℃), extension at 72 ℃ for 2min, and 35 cycles; ③ keeping the temperature at 72 ℃ for 10 min; and fourthly, storing at 4 ℃. After the PCR product is detected by 1.0% agarose gel electrophoresis, an amplification result (figure 2) is observed under an ultraviolet gel imaging system, and the amplification result shows that the length of the trnG-trnS sequence is about 1600 bp.
4) Cloning of PCR products:
and (3) recovering the target band by using an AxyPrep DNA gel recovery kit, recovering and detecting by using 1.2% agarose gel electrophoresis, and taking the purified target DNA as a sequencing template. The recovered product was ligated to a T vector (pGEM-T) using pLB zero background rapid cloning kit, and then transformed into E.coli DH 5. alpha. for culture. Wherein, the amplification result is observed under an ultraviolet gel imaging system, and the trnG-trnS gene has good amplification result, clear band and obvious cloning result by taking the 26-numbered lycium barbarum as an example and referring to the attached figure 3 according to the analysis shown in the figure 3.
5) Sequencing and analysis of sequences
After PCR detection is carried out on positive colonies, the colonies containing target fragments are cultured by an LB liquid culture medium, 3 colonies are selected from each material and sent to a company Limited in Biotechnology engineering (Shanghai) for sequencing, and trnG-trnS sequences of 36 samples are obtained.
The obtained DNA barcode gene sequence is subjected to homology comparison with published sequences in an NCBI database by using DNAMAN, and the DNA barcode gene sequences of 36 parts of medlar materials are respectively subjected to alignment arrangement by using a Clustal X program. A phylogenetic analysis software MEGA7.0 is used for obtaining 1673bp of the total length of the trnG-trnG sequence, 1617 conservative sites accounting for 96.6 percent and 30 variant sites accounting for 1.8 percent, wherein 10 information sites and 20 self-descendent sites have the base transition value of 0.7 and the average value of GC content of 32.3 percent.
After sequence comparison and analysis, 10 germplasms of Huangguo, Lycium ruthenicum Murr, Ningnong wolfberry No. 5, HZ-13-01, ZH-13-08, W-12-27, W-11-15, W-13-26, W-12-26 and Changji wolfberry lack 1 base A at the position of 256 bp; transversions occur at 526bp, 592bp and 1029 bp; a switch occurs at 1365 bp; CT is changed to AA at 1592 bp; at 1612bp the TA transversion is converted to AG. Wherein the yellow fruit change, the black fruit medlar and the Changji medlar are inverted at 935bp, and 2 basic groups TT are inserted at 1410bp of the yellow fruit change; 1 base T is inserted into 1410bp of the lycium ruthenicum; changji wolfberry has 1 base A deleted in 72bp, transversion in 1051bp and other 7 germplasm conversion in 1074. HZ-13-01 is converted at 1062 bp; w-12-27 is converted at 1582 bp; w-13-26bp lacks 1 base A at 529bp and is converted at 1093 bp; w-12-26 was switched at 1036bp and deleted for 1 base T at 1409 bp.
6 germplasms of Ningqi No. 4, northern Lycium chinense, round fruit Lycium chinense, red branches, Hebei Lycium chinense and Xinjiang Lycium chinense lack 1 base A at a position of 72 bp; the transition occurs at 1365 bp. 2 bases TT is inserted at 1410bp of Ningqi No. 4; northern medlar is converted at 55bp and is converted at 935 bp; the red branches are converted at 423 bp and 708bp, 1 base T is deleted at 1409bp, and a sequence with the length of 22bp is inserted at 501bp, namely CATTTTAATAGTTGTAATATTT; the Hebei medlar is converted at 241bp and 964bp and is converted at 935 bp.
Chinese wolfberry and Yunnan wolfberry lack 2 basic groups AA at the position of 72 bp; deletion of 1 base A at 252 bp; transversions occur at 935bp, 1029 bp. 1 base T is inserted into the Chinese wolfberry at the position of 181 bp; deletion of 6bp sequence at 464bp, namely TTTGAA; 3 bases AAA are inserted at 592 bp. The Yunnan medlar is converted at 381 bp; the transversion occurs at 592 bp.
The Ningqi No. 2 is converted at 1471bp, Ningxia Huangguo is converted at 1097bp and 1252bp, and the sprawl medlar is converted at 664bp, 935bp and 1365 bp; 1 base T is inserted at 1410 bp; transversions occurred at 744bp, 1603 bp.
Phylogenetic trees were constructed by comparing the distribution of sequence intraspecific and interspecific differences (FIG. 4). the clustering graph of the trnG-trnS barcode sequence is divided into two major branches, the Lycium ruthenicum Murr, the Changji Lycium Chinense Murr and the Huangguo mutation are clustered into one major branch with the self-expansion supporting rate of 97, wherein the Lycium ruthenicum Murr, the Changji Lycium Chinense Murr and the Huangguo mutation are clustered into one branch with the closest relationship. The rest 26 varieties are gathered into one big branch, wherein the Chinese wolfberry and the Yunnan wolfberry are gathered into one branch, and the self-expanding support rate is 85; the Chinese wolfberry in Hebei, the Chinese wolfberry in North and the vine are gathered into one Chinese wolfberry, and the self-expanding support rate is 62; xinjiang wolfberry, Ningqi No. 4, red branches and round fruit wolfberry are gathered into one, the self-expanding support rate is 67, and the rest 17 germplasms are gathered into one, so that the genetic relationship is nearest. The inside of each branch has the self-expanding support rate of more than 60, and the identification result is reliable.
Therefore, the DNA bar code provided by the invention can be applied to the construction of the medlar phylogenetic tree and further used for researching the phylogeny of medlar intra-species and interspecies. Further proves the effectiveness and feasibility of the DNA bar code provided by the invention in the variety identification, classification and phylogenetic research of the medlar.
Meanwhile, the embodiment of the invention constructs a trnG-trnS bar code database based on the bar code trnG-trnS sequence, the database contains the varieties of Lycium ruthenicum Murr, Lycium xanthocarpum Murr, Lycium rotundus, Lycium barbarum and the like, and the varieties of local varieties of Lycium ruthenicum Murr, local varieties of Sichuan, northern, Xinjiang, Yunnan, Hebei and the like, and different medlar sources of hybrid populations, space mutation populations, ploidy populations and the like, and are all the medlar with representative germplasm in the whole country, so that an effective basis can be provided for classification and identification of medlar varieties.
By comparing the trnG-trnS sequence of the sample to be identified with the sequence in the trnG-trnS bar code database of the medlar sample, the species of medlar can be effectively identified, the interspecific relationship of medlar can be determined, and an effective basis is provided for classification and identification of the variety of medlar by determining the interspecific relationship between the medlar to be detected and the medlar in the bar code database.
Experimental example 1 identification of variety of Lycium barbarum by barcode database
1. Sampling
10 to-be-detected medlar samples with the serial numbers of SD18-01, SD 18-02, ZJ 18-03, ZJ18-04, No.1 yellow, large leaf yellow, P1806, 16-23-8-10, 16-18-16-15 and x 2-184 are selected, and compared with the barcodes of the trnG-trnS barcode database of the part of medlar samples in the embodiment 1 for identification. The variety of the medlar can not be identified by a morphological method, and the experimental example adopts a DNA bar code technology for identification.
TABLE 4 number of sample and production area information of fructus Lycii to be tested
DNA extraction and concentration detection, PCR amplification, PCR product cloning, sequence sequencing and analysis were the same as in example 1.
3. Analysis of sequence results
MEGA7.0 software sequence alignment and clustering analysis are carried out, and a phylogenetic tree is constructed by adopting an NJ method as an attached figure 5. the clustering plot of the trnG-trnS barcode sequence was divided into two major branches, and Lycium ruthenicum Murr were clearly identified. The Lycium ruthenicum Murr and the Changji Lycium Chinense have one big branch, are related recently, are the Lycium ruthenicum Murr, have the self-expansion rate of 100 with other 12 parts of Lycium chinense, and have high reliability.
The samples P1806, 16-23-8-10, 16-18-16-15 to be tested are gathered into a big branch with Ningqi No.1 (bar code database, number 1) and Ningxia Huangguo (bar code database, number 24), the genetic relationship is nearest, the self-expansion rate is 90, wherein the big branch is gathered into a small branch with 2-184 and 16-23-8-10, and the self-expansion rate between the genetic relationship and other 4 samples is 66. The 4 samples to be tested (P1806, 16-23-8-10, 16-18-16-15 and 2-184) are obtained by crossing the hybridization or hybridization filial generations of the Ningxia wolfberries of different varieties, and genetic distance and cluster analysis show that the genetic relationship between the 4 samples to be tested and the Ningxia wolfberries is nearest.
The sample No.1 yellow and the large leaf yellow to be detected are gathered into one, the genetic relationship is recent, the self-development rate is 100, and the reliability is high. SD18-01, SD 18-02, ZJ 18-03 and ZJ18-04 are gathered into one branch, the genetic relationship is relatively close, the self-development rate is 96, and the reliability is high. Yellow No.1 is the Ningqi No.1 with the property of yellow bud change, and SD18-01, SD 18-02, ZJ 18-03 and ZJ18-04 are Chinese wolfberries in Shandong and Zhejiang areas, so that the branch of the yellow No.1 sample to be detected is closer to the branch of the Ningxia wolfberries and closer in genetic relationship, and is further away from the branches of the SD18-01, SD 18-02, ZJ 18-03 and ZJ 18-04. The self-expansion rate of each branch is more than 60, and the high reliability is realized, so that the samples in different regions can be classified and identified on the basis of the trnG-trnS bar code sequence and the bar code database constructed according to the method.
Genetic distance calculations using MEGA7.0 using the K2P model (Kimura 2-parameter model) are shown in Table 5, where the minimum genetic distance between Huangguo and Lycium ruthenicum Murr, Ningqi No.1 and P1806 is 0.00000, and the maximum genetic distance between Huang No.1 and ZJ18-04 is 0.009192.
TABLE 5 genetic distance analysis of trnG-trnS identification of Lycium varieties
Experimental example 2 identification of Lycium ruthenicum varieties by using barcode database
1. Sampling
8 parts of the medlar samples to be detected with the numbers of B1, B3, B5, B8, B9-1, HB, H5 and 15-1 are selected, and compared with the bar codes of the trnG-trnS bar code database of the part of medlar samples in the embodiment 1 for identification. The variety of the medlar can not be identified by a morphological method, and the experimental example adopts a DNA bar code technology for identification.
TABLE 6 number of sample and production area information of fructus Lycii to be tested
DNA extraction and concentration detection, PCR amplification, PCR product cloning, sequence sequencing and analysis were the same as in example 1.
3. Analysis of sequence results
MEGA7.0 software sequence alignment and clustering analysis are carried out, and a phylogenetic tree is constructed by adopting an NJ method as shown in figure 6. the clustering plot of the trnG-trnS barcode sequence was divided into two major branches, and Lycium ruthenicum Murr were clearly identified. The Ningxia Huangguo No.1 Ningxia wolfberry fruit and Chinese wolfberry fruit are gathered into one piece, and the genetic relationship is recent, the Ningxia Huangguo fruit is red fruit wolfberry fruit, the self-expansion rate of the Ningxia Huangguo fruit and other 9 black fruit wolfberry fruits is 99, and the reliability is high.
8 to-be-detected lycium ruthenicum samples and lycium ruthenicum (bar code database, number 12) are gathered into a large branch, the self-expansion rate is 77, and the reliability is high. Wherein the samples B3 to be detected and 15-1 are gathered into one sample, the genetic relationship between the two samples is nearest, and the self-expansion rate is 55; the number of B8, B1 and H5 is one, the genetic relationship of the three is nearest, the self-development rate is 92, and the reliability is high. The trnG-trnS barcode sequence and the barcode database constructed according to the method can be used for classifying and identifying varieties of lycium ruthenicum samples which cannot be identified according to forms in different areas, but the samples have high similarity and are only used as basic identification.
The genetic distance calculation by using MEGA7.0 and K2P model (Kimura 2-parameter model) is shown in Table 7, the genetic distance between B9-1 and Lycium ruthenicum Murr is 0.000608 minimum, the genetic distance between B8 and Ningxia Huangguo is 0.011048 maximum, and the genetic distance between B8 and Ningxia Huangguo is 0.011048 maximum.
TABLE 7 genetic distance analysis for trnL-trnF identification of Lycium ruthenicum Murr
SEQUENCE LISTING
<110> research institute of Lycium barbarum engineering technology of Ningxia academy of sciences and forestry
<120> DNA bar code for variety identification of medlar and identification method thereof
<130>1
<160>38
<170>PatentIn version 3.5
<210>1
<211>1645
<212>DNA
<213>Ningqi No.1 (L. barbarum Linn)
<400>1
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>2
<211>1645
<212>DNA
<213>Ningqi No.2 (L. barbarum Linn)
<400>2
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tggaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>3
<211>1645
<212>DNA
<213>Ningqi No.3 (L. barbarum Linn)
<400>3
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataatgaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>4
<211>1646
<212>DNA
<213>Ningqi No.4 (L. barbarum Linn)
<400>4
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa atcaagtcat caacggaaag agagggattc gaaccctcgg tacgattaac 120
tcgtacaacg gattagcaat ccgccgcttt agtccactca gccatctctc ccaattgaaa 180
aagataatta ctacatgaga tagcacataa gataaaggaa agaatctttc tttctctctt 240
ttcttcttta ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
tcagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca ttttaattga atagttaata ttcaagcaac aagaaaaaat 540
tcccattttt gctaaaagta aaaaaaatat atatatgaaa tagaaaattc gatcaaaata 600
aaagtctcat ttctctttct gcttttttat gtttaccatc ttgctggact aaaaaaaaga 660
agctttcgag tattccacaa tgcattttta tgttatgatt ttagtggttt tgacgaccct 720
atcttatcct atcttgatta ccacaattcc cctgttcgac aaaagttgca tttgtataca 780
ataatcgaat tgtagcgggt atagtttagt ggtaaaagtg tgattcgttc tattatccct 840
taaatagtta aagggtcctt cggtttgatt cgtattccga tcaaaaactt tatttcttaa 900
aaggattaaa tccttttcct ctcaatgaca gattcgagaa caaatacaca ttctcgtgat 960
ttgtatccaa aggtcactta gacattgaaa aattggatta ttaaattgcg aaacataatt 1020
tttgaattgg atcaatactt ccaattgaat aagtatgaat aaaggatcca tggatgaaga 1080
tagaaagttg atttctaatc gtaactaaat cttcaatttc ttatttgtaa agaagaaagt 1140
gaagcaaaat agctattaaa cgatgacttt ggtttactag agacatcaac atattgtttt 1200
agctcggtgg aaacaaaatc cttttcctca ggatcctatt aaatagaaat agagaacgaa 1260
ataactagaa aggttgttag aatccccctc ttctagaagg atcatctaca aagctattcg 1320
ttttatctgt attcagacca aaagctgaca tagatgttat gggtagaatt cttttttttt 1380
ttttctaatt ttgttcacat cttagatcta taaattgact catctccata aaggagccga 1440
atgaaaccaa agtttcatgt tcggttttga attagagacg ttcaaaataa tgaatcgacg 1500
tcgactataa cccctagcct tccaagctaa cgatgcgggt tcgattcccg ctacccgctc 1560
tctatctatt tattctaaat attttaatct tttcattaaa tcaaatttag tttattagta 1620
ttagtacatc attgaatata caattc 1646
<210>5
<211>1645
<212>DNA
<213>Ningqi No.5 (L. barbarum Linn)
<400>5
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>6
<211>1644
<212>DNA
<213>Ningqi No.6 (L. barbarum Linn)
<400>6
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
ttctaatttt gttcacatct tagatctata aattgactca tctccataaa ggagccgaat 1440
gaaaccaaag tttcatgttc ggttttgaat tagagacgtt caaaataatg aatcgacgtc 1500
gactataacc cctagccttc caagctaacg atgcgggttc gattcccgct acccgctctc 1560
tatctattta ttctaaatat tttaatcttt tcattaaatc aaatttagtt tattagtatt 1620
agtacatcat tgaatataca attc 1644
<210>7
<211>1645
<212>DNA
<213>Ningqi No.7 (L. barbarum Linn)
<400>7
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>8
<211>1645
<212>DNA
<213>L. barbarum Linn
<400>8
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>9
<211>1646
<212>DNA
<213>L. barbarum Linn. var. auranticarpum K.F.Ching var. nov.
<400>9
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
tcagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca tattaattga atagttaata ttcaagcaac aagaaaaaat 540
tcccattttt gctaaaagta aaaaaaaaat atatatgaaa tagaaaattc gatcaaaata 600
aaagtctcat ttctctttct gcttttttat gtttaccatc ttgctggact aaaaaaaaga 660
agctttcgag tattccacaa tgcattttta tgttatgatt ttagtggttt tgacgaccct 720
atcttatcct atcttgatta ccacaattcc cctgttcgac aaaagttgca tttgtataca 780
ataatcgaat tgtagcgggt atagtttagt ggtaaaagtg tgattcgttc tattatccct 840
taaatagtta aagggtcctt cggtttgatt cgtattccga tcaaaaactt tatttcttaa 900
aaggatttaa tccttttcct ctcaatgaca gattcgagaa caaatacaca ttctcgtgat 960
ttgtatccaa aggtcactta gacattgaaa aattggatta tgaaattgcg aaacataatt 1020
tttgaattgg atcaatactt ccaattgaat aagtatgaat aaaggatcca tggatgaaga 1080
tagaaagttg atttctaatc gtaactaaat cttcaatttc ttatttgtaa agaagaaagt 1140
gaagcaaaat agctattaaa cgatgacttt ggtttactag agacatcaac atattgtttt 1200
agctcggtgg aaacaaaatc cttttcctca ggatcctatt aaatagaaat agagaacgaa 1260
ataactagaa aggttgttag aatccccctc ttctagaagg atcatctaca aagctattcg 1320
ttttatctgt attcagacca aaagctgaca tagatgttat gggtagaatt cttttttttt 1380
ttttctaatt ttgttcacat cttagatcta taaattgact catctccata aaggagccga 1440
atgaaaccaa agtttcatgt tcggttttga attagagacg ttcaaaataa tgaatcgacg 1500
tcgactataa cccctagcct tccaagctaa cgatgcgggt tcgattcccg ctacccgctc 1560
tctaaatatt tattctaaat atttagatct tttcattaaa tcaaatttag tttattagta 1620
ttagtacatc attgaatata caattc 1646
<210>10
<211>1645
<212>DNA
<213>Mengqi 1（L. barbarum）
<400>10
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>11
<211>1645
<212>DNA
<213>Lycium barbarum Bianguo
<400>11
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>12
<211>1645
<212>DNA
<213>Lycium ruthenicum Murr.
<400>12
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
tcagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca tattaattga atagttaata ttcaagcaac aagaaaaaat 540
tcccattttt gctaaaagta aaaaaaaaat atatatgaaa tagaaaattc gatcaaaata 600
aaagtctcat ttctctttct gcttttttat gtttaccatc ttgctggact aaaaaaaaga 660
agctttcgag tattccacaa tgcattttta tgttatgatt ttagtggttt tgacgaccct 720
atcttatcct atcttgatta ccacaattcc cctgttcgac aaaagttgca tttgtataca 780
ataatcgaat tgtagcgggt atagtttagt ggtaaaagtg tgattcgttc tattatccct 840
taaatagtta aagggtcctt cggtttgatt cgtattccga tcaaaaactt tatttcttaa 900
aaggatttaa tccttttcct ctcaatgaca gattcgagaa caaatacaca ttctcgtgat 960
ttgtatccaa aggtcactta gacattgaaa aattggatta tgaaattgcg aaacataatt 1020
tttgaattgg atcaatactt ccaattgaat aagtatgaat aaaggatcca tggatgaaga 1080
tagaaagttg atttctaatc gtaactaaat cttcaatttc ttatttgtaa agaagaaagt 1140
gaagcaaaat agctattaaa cgatgacttt ggtttactag agacatcaac atattgtttt 1200
agctcggtgg aaacaaaatc cttttcctca ggatcctatt aaatagaaat agagaacgaa 1260
ataactagaa aggttgttag aatccccctc ttctagaagg atcatctaca aagctattcg 1320
ttttatctgt attcagacca aaagctgaca tagatgttat gggtagaatt cttttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctaaatattt attctaaata tttagatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>13
<211>1645
<212>DNA
<213>lycium
<400>13
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>14
<211>1644
<212>DNA
<213>L. barbarum Linn
<400>14
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
tcagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca tattaattga atagttaata ttcaagcaac aagaaaaaat 540
tcccattttt gctaaaagta aaaaaaaaat atatatgaaa tagaaaattc gatcaaaata 600
aaagtctcat ttctctttct gcttttttat gtttaccatc ttgctggact aaaaaaaaga 660
agctttcgag tattccacaa tgcattttta tgttatgatt ttagtggttt tgacgaccct 720
atcttatcct atcttgatta ccacaattcc cctgttcgac aaaagttgca tttgtataca 780
ataatcgaat tgtagcgggt atagtttagt ggtaaaagtg tgattcgttc tattatccct 840
taaatagtta aagggtcctt cggtttgatt cgtattccga tcaaaaactt tatttcttaa 900
aaggattaaa tccttttcct ctcaatgaca gattcgagaa caaatacaca ttctcgtgat 960
ttgtatccaa aggtcactta gacattgaaa aattggatta tgaaattgcg aaacataatt 1020
tttgaattgg atcaatactt ccaattaaat aagtatgaat aaaggatcca tggatgaaga 1080
tagaaagttg atttctaatc gtaactaaat cttcaatttc ttatttgtaa agaagaaagt 1140
gaagcaaaat agctattaaa cgatgacttt ggtttactag agacatcaac atattgtttt 1200
agctcggtgg aaacaaaatc cttttcctca ggatcctatt aaatagaaat agagaacgaa 1260
ataactagaa aggttgttag aatccccctc ttctagaagg atcatctaca aagctattcg 1320
ttttatctgt attcagacca aaagctgaca tagatgttat gggtagaatt cttttttttt 1380
ttctaatttt gttcacatct tagatctata aattgactca tctccataaa ggagccgaat 1440
gaaaccaaag tttcatgttc ggttttgaat tagagacgtt caaaataatg aatcgacgtc 1500
gactataacc cctagccttc caagctaacg atgcgggttc gattcccgct acccgctctc 1560
taaatattta ttctaaatat ttagatcttt tcattaaatc aaatttagtt tattagtatt 1620
agtacatcat tgaatataca attc 1644
<210>15
<211>1644
<212>DNA
<213>lycium
<400>15
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
tcagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca tattaattga atagttaata ttcaagcaac aagaaaaaat 540
tcccattttt gctaaaagta aaaaaaaaat atatatgaaa tagaaaattc gatcaaaata 600
aaagtctcat ttctctttct gcttttttat gtttaccatc ttgctggact aaaaaaaaga 660
agctttcgag tattccacaa tgcattttta tgttatgatt ttagtggttt tgacgaccct 720
atcttatcct atcttgatta ccacaattcc cctgttcgac aaaagttgca tttgtataca 780
ataatcgaat tgtagcgggt atagtttagt ggtaaaagtg tgattcgttc tattatccct 840
taaatagtta aagggtcctt cggtttgatt cgtattccga tcaaaaactt tatttcttaa 900
aaggattaaa tccttttcct ctcaatgaca gattcgagaa caaatacaca ttctcgtgat 960
ttgtatccaa aggtcactta gacattgaaa aattggatta tgaaattgcg aaacataatt 1020
tttgaattgg atcagtactt ccaattaaat aagtatgaat aaaggatcca tggatgaaga 1080
tagaaagttg atttctaatc gtaactaaat cttcaatttc ttatttgtaa agaagaaagt 1140
gaagcaaaat agctattaaa cgatgacttt ggtttactag agacatcaac atattgtttt 1200
agctcggtgg aaacaaaatc cttttcctca ggatcctatt aaatagaaat agagaacgaa 1260
ataactagaa aggttgttag aatccccctc ttctagaagg atcatctaca aagctattcg 1320
ttttatctgt attcagacca aaagctgaca tagatgttat gggtagaatt cttttttttt 1380
ttctaatttt gttcacatct tagatctata aattgactca tctccataaa ggagccgaat 1440
gaaaccaaag tttcatgttc ggttttgaat tagagacgtt caaaataatg aatcgacgtc 1500
gactataacc cctagccttc caagctaacg atgcgggttc gattcccgct acccgctctc 1560
taaatattta ttctaaatat ttagatcttt tcattaaatc aaatttagtt tattagtatt 1620
agtacatcat tgaatataca attc 1644
<210>16
<211>1644
<212>DNA
<213>lycium
<400>16
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
tcagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca tattaattga atagttaata ttcaagcaac aagaaaaaat 540
tcccattttt gctaaaagta aaaaaaaaat atatatgaaa tagaaaattc gatcaaaata 600
aaagtctcat ttctctttct gcttttttat gtttaccatc ttgctggact aaaaaaaaga 660
agctttcgag tattccacaa tgcattttta tgttatgatt ttagtggttt tgacgaccct 720
atcttatcct atcttgatta ccacaattcc cctgttcgac aaaagttgca tttgtataca 780
ataatcgaat tgtagcgggt atagtttagt ggtaaaagtg tgattcgttc tattatccct 840
taaatagtta aagggtcctt cggtttgatt cgtattccga tcaaaaactt tatttcttaa 900
aaggattaaa tccttttcct ctcaatgaca gattcgagaa caaatacaca ttctcgtgat 960
ttgtatccaa aggtcactta gacattgaaa aattggatta tgaaattgcg aaacataatt 1020
tttgaattgg atcaatactt ccaattaaat aagtatgaat aaaggatcca tggatgaaga 1080
tagaaagttg atttctaatc gtaactaaat cttcaatttc ttatttgtaa agaagaaagt 1140
gaagcaaaat agctattaaa cgatgacttt ggtttactag agacatcaac atattgtttt 1200
agctcggtgg aaacaaaatc cttttcctca ggatcctatt aaatagaaat agagaacgaa 1260
ataactagaa aggttgttag aatccccctc ttctagaagg atcatctaca aagctattcg 1320
ttttatctgt attcagacca aaagctgaca tagatgttat gggtagaatt cttttttttt 1380
ttctaatttt gttcacatct tagatctata aattgactca tctccataaa ggagccgaat 1440
gaaaccaaag tttcatgttc ggttttgaat tagagacgtt caaaataatg aatcgacgtc 1500
gactataacc cctagccttc caagctaacg atgcgggttc gattcccgct acccgctctc 1560
taaatattta ttctaaatat ttagatcttt tcattaaatc aaatttagtt tattagtatt 1620
agtacatcat tgaatataca attc 1644
<210>17
<211>1644
<212>DNA
<213>lycium
<400>17
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
tcagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca tattaattga atagttaata ttcaagcaac aagaaaaaat 540
tcccattttt gctaaaagta aaaaaaaaat atatatgaaa tagaaaattc gatcaaaata 600
aaagtctcat ttctctttct gcttttttat gtttaccatc ttgctggact aaaaaaaaga 660
agctttcgag tattccacaa tgcattttta tgttatgatt ttagtggttt tgacgaccct 720
atcttatcct atcttgatta ccacaattcc cctgttcgac aaaagttgca tttgtataca 780
ataatcgaat tgtagcgggt atagtttagt ggtaaaagtg tgattcgttc tattatccct 840
taaatagtta aagggtcctt cggtttgatt cgtattccga tcaaaaactt tatttcttaa 900
aaggattaaa tccttttcct ctcaatgaca gattcgagaa caaatacaca ttctcgtgat 960
ttgtatccaa aggtcactta gacattgaaa aattggatta tgaaattgcg aaacataatt 1020
tttgaattgg atcaatactt ccaattaaat aagtatgaat aaaggatcca tggatgaaga 1080
tagaaagttg atttctaatc gtaactaaat cttcaatttc ttatttgtaa agaagaaagt 1140
gaagcaaaat agctattaaa cgatgacttt ggtttactag agacatcaac atattgtttt 1200
agctcggtgg aaacaaaatc cttttcctca ggatcctatt aaatagaaat agagaacgaa 1260
ataactagaa aggttgttag aatccccctc ttctagaagg atcatctaca aagctattcg 1320
ttttatctgt attcagacca aaagctgaca tagatgttat gggtagaatt cttttttttt 1380
ttctaatttt gttcacatct tagatctata aattgactca tctccataaa ggagccgaat 1440
gaaaccaaag tttcatgttc ggttttgaat tagagacgtt caaaataatg aatcgacgtc 1500
gactataacc cctagccttc caagctaacg atgcgggttc gattcccgct actcgctctc 1560
taaatattta ttctaaatat ttagatcttt tcattaaatc aaatttagtt tattagtatt 1620
agtacatcat tgaatataca attc 1644
<210>18
<211>1644
<212>DNA
<213>lycium
<400>18
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
tcagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca tattaattga atagttaata ttcaagcaac aagaaaaaat 540
tcccattttt gctaaaagta aaaaaaaaat atatatgaaa tagaaaattc gatcaaaata 600
aaagtctcat ttctctttct gcttttttat gtttaccatc ttgctggact aaaaaaaaga 660
agctttcgag tattccacaa tgcattttta tgttatgatt ttagtggttt tgacgaccct 720
atcttatcct atcttgatta ccacaattcc cctgttcgac aaaagttgca tttgtataca 780
ataatcgaat tgtagcgggt atagtttagt ggtaaaagtg tgattcgttc tattatccct 840
taaatagtta aagggtcctt cggtttgatt cgtattccga tcaaaaactt tatttcttaa 900
aaggattaaa tccttttcct ctcaatgaca gattcgagaa caaatacaca ttctcgtgat 960
ttgtatccaa aggtcactta gacattgaaa aattggatta tgaaattgcg aaacataatt 1020
tttgaattgg atcaatactt ccaattaaat aagtatgaat aaaggatcca tggatgaaga 1080
tagaaagttg atttctaatc gtaactaaat cttcaatttc ttatttgtaa agaagaaagt 1140
gaagcaaaat agctattaaa cgatgacttt ggtttactag agacatcaac atattgtttt 1200
agctcggtgg aaacaaaatc cttttcctca ggatcctatt aaatagaaat agagaacgaa 1260
ataactagaa aggttgttag aatccccctc ttctagaagg atcatctaca aagctattcg 1320
ttttatctgt attcagacca aaagctgaca tagatgttat gggtagaatt cttttttttt 1380
ttctaatttt gttcacatct tagatctata aattgactca tctccataaa ggagccgaat 1440
gaaaccaaag tttcatgttc ggttttgaat tagagacgtt caaaataatg aatcgacgtc 1500
gactataacc cctagccttc caagctaacg atgcgggttc gattcccgct acccgctctc 1560
taaatattta ttctaaatat ttagatcttt tcattaaatc aaatttagtt tattagtatt 1620
agtacatcat tgaatataca attc 1644
<210>19
<211>1643
<212>DNA
<213>lycium
<400>19
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
tcagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca tattaattga atagttaata ttcaagcaac aagaaaaaat 540
tcccattttt gctaaaagta aaaaaaaata tatatgaaat agaaaattcg atcaaaataa 600
aagtctcatt tctctttctg cttttttatg tttaccatct tgctggacta aaaaaaagaa 660
gctttcgagt attccacaat gcatttttat gttatgattt tagtggtttt gacgacccta 720
tcttatccta tcttgattac cacaattccc ctgttcgaca aaagttgcat ttgtatacaa 780
taatcgaatt gtagcgggta tagtttagtg gtaaaagtgt gattcgttct attatccctt 840
aaatagttaa agggtccttc ggtttgattc gtattccgat caaaaacttt atttcttaaa 900
aggattaaat ccttttcctc tcaatgacag attcgagaac aaatacacat tctcgtgatt 960
tgtatccaaa ggtcacttag acattgaaaa attggattat gaaattgcga aacataattt 1020
ttgaattgga tcaatacttc caattaaata agtatgaata aagggtccat ggatgaagat 1080
agaaagttga tttctaatcg taactaaatc ttcaatttct tatttgtaaa gaagaaagtg 1140
aagcaaaata gctattaaac gatgactttg gtttactaga gacatcaaca tattgtttta 1200
gctcggtgga aacaaaatcc ttttcctcag gatcctatta aatagaaata gagaacgaaa 1260
taactagaaa ggttgttaga atccccctct tctagaagga tcatctacaa agctattcgt 1320
tttatctgta ttcagaccaa aagctgacat agatgttatg ggtagaattc tttttttttt 1380
tctaattttg ttcacatctt agatctataa attgactcat ctccataaag gagccgaatg 1440
aaaccaaagt ttcatgttcg gttttgaatt agagacgttc aaaataatga atcgacgtcg 1500
actataaccc ctagccttcc aagctaacga tgcgggttcg attcccgcta cccgctctct 1560
aaatatttat tctaaatatt tagatctttt cattaaatca aatttagttt attagtatta 1620
gtacatcatt gaatatacaa ttc 1643
<210>20
<211>1643
<212>DNA
<213>lycium
<400>20
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cggaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
tcagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca tattaattga atagttaata ttcaagcaac aagaaaaaat 540
tcccattttt gctaaaagta aaaaaaaaat atatatgaaa tagaaaattc gatcaaaata 600
aaagtctcat ttctctttct gcttttttat gtttaccatc ttgctggact aaaaaaaaga 660
agctttcgag tattccacaa tgcattttta tgttatgatt ttagtggttt tgacgaccct 720
atcttatcct atcttgatta ccacaattcc cctgttcgac aaaagttgca tttgtataca 780
ataatcgaat tgtagcgggt atagtttagt ggtaaaagtg tgattcgttc tattatccct 840
taaatagtta aagggtcctt cggtttgatt cgtattccga tcaaaaactt tatttcttaa 900
aaggattaaa tccttttcct ctcaatgaca gattcgagaa caaatacaca ttctcgtgat 960
ttgtatccaa aggtcactta gacattgaaa aattggatta tgaaattgcg aaacataatt 1020
tttgaattgg atcaatactt ccaattaaat aagtatgaat aaaggatcca tggatgaaga 1080
tagaaagttg atttctaatc gtaactaaat cttcaatttc ttatttgtaa agaagaaagt 1140
gaagcaaaat agctattaaa cgatgacttt ggtttactag agacatcaac atattgtttt 1200
agctcggtgg aaacaaaatc cttttcctca ggatcctatt aaatagaaat agagaacgaa 1260
ataactagaa aggttgttag aatccccctc ttctagaagg atcatctaca aagctattcg 1320
ttttatctgt attcagacca aaagctgaca tagatgttat gggtagaatt cttttttttt 1380
tctaattttg ttcacatctt agatctataa attgactcat ctccataaag gagccgaatg 1440
aaaccaaagt ttcatgttcg gttttgaatt agagacgttc aaaataatga atcgacgtcg 1500
actataaccc ctagccttcc aagctaacga tgcgggttcg attcccgcta cccgctctct 1560
aaatatttat tctaaatatt tagatctttt cattaaacca aatttagttt attagtatta 1620
gtacatcatt gaatatacaa ttc 1643
<210>21
<211>1645
<212>DNA
<213>Lycium chinense MilL. var. potaninii (Pojark.) A. M. Lu
<400>21
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcggaaatt 60
gaaaaaaaaa atcaagtcat caacggaaag agagggattc gaaccctcgg tacgattaac 120
tcgtacaacg gattagcaat ccgccgcttt agtccactca gccatctctc ccaattgaaa 180
aagataatta ctacatgaga tagcacataa gataaaggaa agaatctttc tttctctctt 240
ttcttcttta ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
tcagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca ttttaattga atagttaata ttcaagcaac aagaaaaaat 540
tcccattttt gctaaaagta aaaaaaatat atatatgaaa tagaaaattc gatcaaaata 600
aaagtctcat ttctctttct gcttttttat gtttaccatc ttgctggact aaaaaaaaga 660
agctttcgag tattccacaa tgcattttta tgttatgatt ttagtggttt tgacgaccct 720
atcttatcct atcttgatta ccacaattcc cctgttcgac aaaagttgca tttgtataca 780
ataatcgaat tgtagcgggt atagtttagt ggtaaaagtg tgattcgttc tattatccct 840
taaatagtta aagggtcctt cggtttgatt cgtattccga tcaaaaactt tatttcttaa 900
aaggatttaa tccttttcct ctcaatgaca gattcgagaa caaatacaca ttctcgtgat 960
ttgtatccaa aggtcactta gacattgaaa aattggatta ttaaattgcg aaacataatt 1020
tttgaattgg atcaatactt ccaattgaat aagtatgaat aaaggatcca tggatgaaga 1080
tagaaagttg atttctaatc gtaactaaat cttcaatttc ttatttgtaa agaagaaagt 1140
gaagcaaaat agctattaaa cgatgacttt ggtttactag agacatcaac atattgtttt 1200
agctcggtgg aaacaaaatc cttttcctca ggatcctatt aaatagaaat agagaacgaa 1260
ataactagaa aggttgttag aatccccctc ttctagaagg atcatctaca aagctattcg 1320
ttttatctgt attcagacca aaagctgaca tagatgttat gggtagaatt cttttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>22
<211>1645
<212>DNA
<213>lycium
<400>22
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa atcaagtcat caacggaaag agagggattc gaaccctcgg tacgattaac 120
tcgtacaacg gattagcaat ccgccgcttt agtccactca gccatctctc ccaattgaaa 180
aagataatta ctacatgaga tagcacataa gataaaggaa agaatctttc tttctctctt 240
ttcttcttta ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
tcagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca ttttaattga atagttaata ttcaagcaac aagaaaaaat 540
tcccattttt gctaaaagta aaaaaaatat atatatgaaa tagaaaattc gatcaaaata 600
aaagtctcat ttctctttct gcttttttat gtttaccatc ttgctggact aaaaaaaaga 660
agctttcgag tattccacaa tgcattttta tgttatgatt ttagtggttt tgacgaccct 720
atcttatcct atcttgatta ccacaattcc cctgttcgac aaaagttgca tttgtataca 780
ataatcgaat tgtagcgggt atagtttagt ggtaaaagtg tgattcgttc tattatccct 840
taaatagtta aagggtcctt cggtttgatt cgtattccga tcaaaaactt tatttcttaa 900
aaggattaaa tccttttcct ctcaatgaca gattcgagaa caaatacaca ttctcgtgat 960
ttgtatccaa aggtcactta gacattgaaa aattggatta ttaaattgcg aaacataatt 1020
tttgaattgg atcaatactt ccaattgaat aagtatgaat aaaggatcca tggatgaaga 1080
tagaaagttg atttctaatc gtaactaaat cttcaatttc ttatttgtaa agaagaaagt 1140
gaagcaaaat agctattaaa cgatgacttt ggtttactag agacatcaac atattgtttt 1200
agctcggtgg aaacaaaatc cttttcctca ggatcctatt aaatagaaat agagaacgaa 1260
ataactagaa aggttgttag aatccccctc ttctagaagg atcatctaca aagctattcg 1320
ttttatctgt attcagacca aaagctgaca tagatgttat gggtagaatt cttttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>23
<211>1645
<212>DNA
<213>lycium
<400>23
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>24
<211>1645
<212>DNA
<213>lycium
<400>24
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaaaggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc gtggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat cctttccctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>25
<211>1645
<212>DNA
<213>Lycium chinense
<400>25
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>26
<211>1645
<212>DNA
<213>Damaye (L. barbarum Linn)
<400>26
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>27
<211>1645
<212>DNA
<213>Baihua(L. barbarum)
<400>27
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>28
<211>1625
<212>DNA
<213>L. chinense Mill. var.
<400>28
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa tcaagtcatc aacggaaaga gagggattcg aaccctcggt acgattaact 120
cgtacaacgg attagcaatc cgccgcttta gtccactcag ccatctctcc caattgaata 180
aagataatta ctacatgaga tagcacataa gataaaggaa agaatctttc tttctctctt 240
ttcttctttc tatattatat agatatgtac aacttttatc atcaatttcc tttatctctt 300
tatctaaagt aaaggaaggg ctcagaagag ccaagaatat caagaaaaat aaagaagacc 360
gcttttcttt gtcttgattt tgttcgaaag gaccctctta ttctcatggc ctggtctggt 420
cagtacccag ccgggcctct tttgttccaa cgaatttgaa aacaaaaatg cctgttatag 480
ttgtaatatt tcattttaat tgaatagtta atattcaagc aacaagaaaa aattcccatt 540
tttgctaaaa gtaaaaaaaa aatatatata tgaaatagaa aattcgatca aaataaaagt 600
ctcatttctc tttctgcttt tttatgttta ccatcttgct ggactaaaaa aaagaagctt 660
tcgagtattc cacaatgcat ttttatgtta tgattttagt ggttttgacg accctatctt 720
atcctatctt gattaccaca attcccctgt tcgacaaaag ttgcatttgt atacaataat 780
cgaattgtag cgggtatagt ttagtggtaa aagtgtgatt cgttctatta tcccttaaat 840
agttaaaggg tccttcggtt tgattcgtat tccgatcaaa aactttattt cttaaaagga 900
tttaatcctt ttcctctcaa tgacagattc gagaacaaat acacattctc gtgatttgta 960
tccaaaggtc acttagacat tgaaaaattg gattatgaaa ttgcgaaaca taatttttga 1020
attggatcaa tacttccaat tgaataagta tgaataaagg atccatggat gaagatagaa 1080
agttgatttc taatcgtaac taaatcttca atttcttatt tgtaaagaag aaagtgaagc 1140
aaaatagcta ttaaacgatg actttggttt actagagaca tcaacatatt gttttagctc 1200
ggtggaaaca aaatcctttt cctcaggatc ctattaaata gaaatagaga acgaaataac 1260
tagaaaggtt gttagaatcc ccctcttcta gaaggatcat ctacaaagct attcgtttta 1320
tctgtattca gaccaaaagc tgacatagat gttatgggta gaattctttt tttttttcta 1380
attttgttca catcttagat ctataaattg actcatctcc ataaaggagc cgaatgaaac 1440
caaagtttca tgttcggttt tgaattagag acgttcaaaa taatgaatcg acgtcgacta 1500
taacccctag ccttccaagc taacgatgcg ggttcgattc ccgctacccg ctctctatct 1560
atttattcta aatattttaa tcttttcatt aaatcaaatt tagtttatta gtattagtac 1620
atcat 1625
<210>29
<211>1643
<212>DNA
<213>Lycium yunnanenseKuang et A.M.Lu
<400>29
ttaggatttg gtctattccacacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa tcaagtcatc aacggaaaga gagggattcg aaccctcggt acgattaact 120
cgtacaacgg attagcaatc cgccgcttta gtccactcag ccatctctcc caattgaaaa 180
agataattac tacatgagat agcacataag ataaaggaaa gaatctttct ttctctcttt 240
tcttctttct atattatata gatatgtaca acttttatca tcaatttcct ttatctcttt 300
atctaaagta aagaaagggc tcagaagagc caagaatatc aagaaaaata aagaagaccg 360
cttttctttg tcttgatttt gttcgaaagg accctcttat tctcatggcc tggtctggtc 420
agtacccagc cgggcctctt ttgttccaac gaatttgaat ttgaaaacaa aaatgcctgt 480
tatagttgta atatttcatt ttaattgaat agttaatatt caagcaacaa gaaaaaattc 540
ccatttttgc taaaagtaaa aaaaaaatat atatgaaata gaaaattcga tcaaaataaa 600
agtctcattt ctctttctgc ttttttatgt ttaccatctt gctggactaa aaaaaagaag 660
ctttcgagta ttccacaatg catttttatg ttatgatttt agtggttttg acgaccctat 720
cttatcctat cttgattacc acaattcccc tgttcgacaa aagttgcatt tgtatacaat 780
aatcgaattg tagcgggtat agtttagtgg taaaagtgtg attcgttcta ttatccctta 840
aatagttaaa gggtccttcg gtttgattcg tattccgatc aaaaacttta tttcttaaaa 900
ggatttaatc cttttcctct caatgacaga ttcgagaaca aatacacatt ctcgtgattt 960
gtatccaaag gtcacttaga cattgaaaaa ttggattatg aaattgcgaa acataatttt 1020
tgaattggat caatacttcc aattgaataa gtatgaataa aggatccatg gatgaagata 1080
gaaagttgat ttctaatcgt aactaaatct tcaatttctt atttgtaaag aagaaagtga 1140
agcaaaatag ctattaaacg atgactttgg tttactagag acatcaacat attgttttag 1200
ctcggtggaa acaaaatcct tttcctcagg atcctattaa atagaaatag agaacgaaat 1260
aactagaaag gttgttagaa tccccctctt ctagaaggat catctacaaa gctattcgtt 1320
ttatctgtat tcagaccaaa agctgacata gatgttatgg gtagaattct tttttttttt 1380
tctaattttg ttcacatctt agatctataa attgactcat ctccataaag gagccgaatg 1440
aaaccaaagt ttcatgttcg gttttgaatt agagacgttc aaaataatga atcgacgtcg 1500
actataaccc ctagccttcc aagctaacga tgcgggttcg attcccgcta cccgctctct 1560
atctatttat tctaaatatt ttaatctttt cattaaatca aatttagttt attagtatta 1620
gtacatcatt gaatatacaa ttc 1643
<210>30
<211>1646
<212>DNA
<213>Manshenggouqi (L.bararum)
<400>30
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttactat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaacc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattta atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagacc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
ttttataatt ttgttcacat cttagatcta taaattgact catctccata aaggagccga 1440
atgaaaccaa agtttcatgt tcggttttga attagagacg ttcaaaataa tgaatcgacg 1500
tcgactataa cccctagcct tccaagctaa cgatgcgggt tcgattcccg ctacccgctc 1560
tctatctatt tattcgaaat attttaatct tttcattaaa tcaaatttag tttattagta 1620
ttagtacatc attgaatata caattc 1646
<210>31
<211>1645
<212>DNA
<213>Ziguogouqi (L.barbarum)
<400>31
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>32
<211>1664
<212>DNA
<213>Lycium dasystemum
<400>32
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa atcaagtcat caacggaaag agagggattc gaaccctcgg tacgattaac 120
tcgtacaacg gattagcaat ccgccgcttt agtccactca gccatctctc ccaattgaaa 180
aagataatta ctacatgaga tagcacataa gataaaggaa agaatctttc tttctctctt 240
ttcttcttta ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
ccagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca ttttaatagt tgtaatattt cattttaatt gaatagttaa 540
tattcaagca acaagaaaaa attcccattt ttgctaaaag taaaaaaaat atatatatga 600
aatagaaaat tcgatcaaaa taaaagtctc atttctcttt ctgctttttt atgtttacca 660
tcttgctgga ctaaaaaaaa gaagctttcg agtattccac aacgcatttt tatgttatga 720
ttttagtggt tttgacgacc ctatcttatc ctatcttgat taccacaatt cccctgttcg 780
acaaaagttg catttgtata caataatcga attgtagcgg gtatagttta gtggtaaaag 840
tgtgattcgt tctattatcc cttaaatagt taaagggtcc ttcggtttga ttcgtattcc 900
gatcaaaaac tttatttctt aaaaggatta aatccttttc ctctcaatga cagattcgag 960
aacaaataca cattctcgtg atttgtatcc aaaggtcact tagacattga aaaattggat 1020
tattaaattg cgaaacataa tttttgaatt ggatcaatac ttccaattga ataagtatga 1080
ataaaggatc catggatgaa gatagaaagt tgatttctaatcgtaactaa atcttcaatt 1140
tcttatttgt aaagaagaaa gtgaagcaaa atagctatta aacgatgact ttggtttact 1200
agagacatca acatattgtt ttagctcggt ggaaacaaaa tccttttcct caggatccta 1260
ttaaatagaa atagagaacg aaataactag aaaggttgtt agaatccccc tcttctagaa 1320
ggatcatcta caaagctatt cgttttatct gtattcagac caaaagctga catagatgtt 1380
atgggtagaa ttcttttttt ttctaatttt gttcacatct tagatctata aattgactca 1440
tctccataaa ggagccgaat gaaaccaaag tttcatgttc ggttttgaat tagagacgtt 1500
caaaataatg aatcgacgtc gactataacc cctagccttc caagctaacg atgcgggttc 1560
gattcccgct acccgctctc tatctattta ttctaaatat tttaatcttt tcattaaatc 1620
aaatttagtt tattagtatt agtacatcat tgaatataca attc 1664
<210>33
<211>1645
<212>DNA
<213>lycium
<400>33
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa atcaagtcat caacggaaag agagggattc gaaccctcgg tacgattaac 120
tcgtacaacg gattagcaat ccgccgcttt agtccactca gccatctctc ccaattgaaa 180
aagataatta ctacatgaga tagcacataa gataaaggaa agaatctttc tttctctcct 240
ttcttcttta ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
tcagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca ttttaattga atagttaata ttcaagcaac aagaaaaaat 540
tcccattttt gctaaaagta aaaaaaatat atatatgaaa tagaaaattc gatcaaaata 600
aaagtctcat ttctctttct gcttttttat gtttaccatc ttgctggact aaaaaaaaga 660
agctttcgag tattccacaa tgcattttta tgttatgatt ttagtggttt tgacgaccct 720
atcttatcct atcttgatta ccacaattcc cctgttcgac aaaagttgca tttgtataca 780
ataatcgaat tgtagcgggt atagtttagt ggtaaaagtg tgattcgttc tattatccct 840
taaatagtta aagggtcctt cggtttgatt cgtattccga tcaaaaactt tatttcttaa 900
aaggatttaa tccttttcct ctcaatgaca gattcgggaa caaatacaca ttctcgtgat 960
ttgtatccaa aggtcactta gacattgaaa aattggatta ttaaattgcg aaacataatt 1020
tttgaattgg atcaatactt ccaattgaat aagtatgaat aaaggatcca tggatgaaga 1080
tagaaagttg atttctaatc gtaactaaat cttcaatttc ttatttgtaa agaagaaagt 1140
gaagcaaaat agctattaaa cgatgacttt ggtttactag agacatcaac atattgtttt 1200
agctcggtgg aaacaaaatc cttttcctca ggatcctatt aaatagaaat agagaacgaa 1260
ataactagaa aggttgttag aatccccctc ttctagaagg atcatctaca aagctattcg 1320
ttttatctgt attcagacca aaagctgaca tagatgttat gggtagaatt cttttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>34
<211>1645
<212>DNA
<213>Xiaomaye (L. barbarum Linn)
<400>34
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa aatcaagtca tcaacggaaa gagagggatt cgaaccctcg gtacgattaa 120
ctcgtacaac ggattagcaa tccgccgctt tagtccactc agccatctct cccaattgaa 180
aaagataatt actacatgag atagcacata agataaagga aagaatcttt ctttctctct 240
tttcttcttt actatattat atagatatgt acaactttta tcatcaattt cctttatctc 300
tttatctaaa gtaaaggaag ggctcagaag agccaagaat atcaagaaaa ataaagaaga 360
ccgcttttct ttgtcttgat tttgttcgaa aggaccctct tattctcatg gcctggtctg 420
gtcagtaccc agccgggcct cttttgttcc aacgaatttg aatttgaaaa caaaaatgcc 480
tgttatagtt gtaatatttc attttaattg aatagttaat attcaagcaa caagaaaaaa 540
ttcccatttt tgctaaaagt aaaaaaaata tatatatgaa atagaaaatt cgatcaaaat 600
aaaagtctca tttctctttc tgctttttta tgtttaccat cttgctggac taaaaaaaag 660
aagctttcga gtattccaca atgcattttt atgttatgat tttagtggtt ttgacgaccc 720
tatcttatcc tatcttgatt accacaattc ccctgttcga caaaagttgc atttgtatac 780
aataatcgaa ttgtagcggg tatagtttag tggtaaaagt gtgattcgtt ctattatccc 840
ttaaatagtt aaagggtcct tcggtttgat tcgtattccg atcaaaaact ttatttctta 900
aaaggattaa atccttttcc tctcaatgac agattcgaga acaaatacac attctcgtga 960
tttgtatcca aaggtcactt agacattgaa aaattggatt attaaattgc gaaacataat 1020
ttttgaattg gatcaatact tccaattgaa taagtatgaa taaaggatcc atggatgaag 1080
atagaaagtt gatttctaat cgtaactaaa tcttcaattt cttatttgta aagaagaaag 1140
tgaagcaaaa tagctattaa acgatgactt tggtttacta gagacatcaa catattgttt 1200
tagctcggtg gaaacaaaat ccttttcctc aggatcctat taaatagaaa tagagaacga 1260
aataactaga aaggttgtta gaatccccct cttctagaag gatcatctac aaagctattc 1320
gttttatctg tattcagatc aaaagctgac atagatgtta tgggtagaat tctttttttt 1380
tttctaattt tgttcacatc ttagatctat aaattgactc atctccataa aggagccgaa 1440
tgaaaccaaa gtttcatgtt cggttttgaa ttagagacgt tcaaaataat gaatcgacgt 1500
cgactataac ccctagcctt ccaagctaac gatgcgggtt cgattcccgc tacccgctct 1560
ctatctattt attctaaata ttttaatctt ttcattaaat caaatttagt ttattagtat 1620
tagtacatca ttgaatatac aattc 1645
<210>35
<211>1643
<212>DNA
<213>lycium
<400>35
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa atcaagtcat caacggaaag agagggattc gaaccctcgg tacgattaac 120
tcgtacaacg gattagcaat ccgccgcttt agtccactca gccatctctc ccaattgaaa 180
aagataatta ctacatgaga tagcacataa gataaaggaa agaatctttc tttctctctt 240
ttcttctttc tatattatat agatatgtac aacttttatc atcaatttcc tttatctctt 300
tatctaaagt aaaggaaggg ctcagaagag ccaagaatat caagaaaaat aaagaagacc 360
gcttttcttt gtcttgattt tgttcgaaag gaccctctta ttctcatggc ctggtctggt 420
cagtacccag ccgggcctct tttgttccaa cgaatttgaa tttgaaaaca aaaatgcctg 480
ttatagttgt aatatttcat attaattgaa tagttaatat tcaagcaaca agaaaaaatt 540
cccatttttg ctaaaagtaa aaaaaaaata tatatgaaat agaaaattcg atcaaaataa 600
aagtctcatt tctctttctg cttttttatg tttaccatct tgctggacta aaaaaaagaa 660
gctttcgagt attccacaat gcatttttat gttatgattt tagtggtttt gacgacccta 720
tcttatccta tcttgattac cacaattccc ctgttcgaca aaagttgcat ttgtatacaa 780
taatcgaatt gtagcgggta tagtttagtg gtaaaagtgt gattcgttct attatccctt 840
aaatagttaa agggtccttc ggtttgattc gtattccgat caaaaacttt atttcttaaa 900
aggatttaat ccttttcctc tcaatgacag attcgagaac aaatacacat tctcgtgatt 960
tgtatccaaa ggtcacttag acattgaaaa attggattat gaaattgcga aacataattt 1020
ttcaattgga tcaatacttc caattgaata agtatgaata aaggatccat ggatgaagat 1080
agaaagttga tttctaatcg taactaaatc ttcaatttct tatttgtaaa gaagaaagtg 1140
aagcaaaata gctattaaac gatgactttg gtttactaga gacatcaaca tattgtttta 1200
gctcggtgga aacaaaatcc ttttcctcag gatcctatta aatagaaata gagaacgaaa 1260
taactagaaa ggttgttaga atccccctct tctagaagga tcatctacaa agctattcgt 1320
tttatctgta ttcagaccaa aagctgacat agatgttatg ggtagaattc tttttttttt 1380
tctaattttg ttcacatctt agatctataa attgactcat ctccataaag gagccgaatg 1440
aaaccaaagt ttcatgttcg gttttgaatt agagacgttc aaaataatga atcgacgtcg 1500
actataaccc ctagccttcc aagctaacga tgcgggttcg attcccgcta cccgctctct 1560
aaatatttat tctaaatatt tagatctttt cattaaatca aatttagttt attagtatta 1620
gtacatcatt gaatatacaa ttc 1643
<210>36
<211>1644
<212>DNA
<213>Lycium dasystemumPojark
<400>36
ttaggatttg gtctattcca cacatttaac taagaataag aacaaaggat ttcgaaaatt 60
gaaaaaaaaa atcaagtcat caacggaaag agagggattc gaaccctcgg tacgattaac 120
tcgtacaacg gattagcaat ccgccgcttt agtccactca gccatctctc ccaattgaaa 180
aagataatta ctacatgaga tagcacataa gataaaggaa agaatctttc tttctctctt 240
ttcttcttta ctatattata tagatatgta caacttttat catcaatttc ctttatctct 300
ttatctaaag taaaggaagg gctcagaaga gccaagaata tcaagaaaaa taaagaagac 360
cgcttttctt tgtcttgatt ttgttcgaaa ggaccctctt attctcatgg cctggtctgg 420
tcagtaccca gccgggcctc ttttgttcca acgaatttga atttgaaaac aaaaatgcct 480
gttatagttg taatatttca ttttaattga atagttaata ttcaagcaac aagaaaaaat 540
tcccattttt gctaaaagta aaaaatatat atatatgaaa tagaaaattc gatcaaaata 600
aaagtctcat ttctctttct gcttttttat gtttaccatc ttgctggact aaaaaaaaga 660
agctttcgag tattccacaa tgcattttta tgttatgatt ttagtggttt tgacgaccct 720
atcttatcct atcttgatta ccacaattcc cctgttcgac aaaagttgca tttgtataca 780
ataatcgaat tgtagcgggt atagtttagt ggtaaaagtg tgattcgttc tattatccct 840
taaatagtta aagggtcctt cggtttgatt cgtattccga tcaaaaactt tatttcttaa 900
aaggattaaa tccttttcct ctcaatgaca gattcgagaa caaatacaca ttctcgtgat 960
ttgtatccaa aggtcactta gacattgaaa aattggatta ttaaattgcg aaacataatt 1020
tttgaattgg atcaatactt ccaattgaat aagtatgaat aaaggatcca tggatgaaga 1080
tagaaagttg atttctaatc gtaactaaat cttcaatttc ttatttgtaa agaagaaagt 1140
gaagcaaaat agctattaaa cgatgacttt ggtttactag agacatcaac atattgtttt 1200
agctcggtgg aaacaaaatc cttttcctca ggatcctatt aaatagaaat agagaacgaa 1260
ataactagaa aggttgttag aatccccctc ttctagaagg atcatctaca aagctattcg 1320
ttttatctgt attcagacca aaagctgaca tagatgttat gggtagaatt cttttttttt 1380
ttctaatttt gttcacatct tagatctata aattgactca tctccataaa ggagccgaat 1440
gaaaccaaag tttcatgttc ggttttgaat tagagacgtt caaaataatg aatcgacgtc 1500
gactataacc cctagccttc caagctaacg atgcgggttc gattcccgct acccgctctc 1560
tatctattta ttctaaatat tttaatcttt tcattaaatc aaatttagtt tattagtatt 1620
agtacatcat tgaatataca attc 1644
<210>37
<211>19
<212>DNA
<213> Artificial sequence
<400>37
<210>38
<211>20
<212>DNA
<213> Artificial sequence
<400>38
Claims (7)
1. A method for identifying Chinese wolfberry varieties by using DNA bar codes is characterized in that: the DNA barcode is trnG-trnS barcode.
2. The method for identifying the variety of lycium barbarum based on DNA barcode according to claim 1, wherein: the method comprises the following steps:
1) extracting genome DNA from a medlar sample;
2) amplifying a sequence fragment of the trnG-trnS bar code by using the genomic DNA as a template and primers shown as nucleotide sequences SEQ ID NO.37 and SEQ ID NO.38 to obtain a PCR product;
3) sequencing the PCR amplification product;
4) and (5) constructing a phylogenetic tree and identifying the medlar.
3. The method for rapidly identifying medlar based on DNA barcodes as claimed in claim 2, which is characterized in that: in the step 1), DNA of the wolfberry sample is extracted by using a Kit, wherein the Kit is a DNA secure Plant Kit.
4. The method for rapidly identifying medlar based on DNA barcodes as claimed in claim 2, which is characterized in that: in the step 2), the reaction system of PCR amplification is as follows: firstly, pre-denaturation is carried out for 2min at 94 ℃; ② denaturation at 94 ℃ for 30s, annealing at 55-60 ℃ for 30s, extension at 72 ℃ for 2min, and 35 cycles; ③ keeping the temperature at 72 ℃ for 10 min; fourthly, preserving at 4 ℃; and carrying out detection verification on the obtained PCR product.
5. A trnG-trnS barcode database of a Lycium barbarum sample constructed by the method of any one of claims 1-4, comprising trnG-trnS barcode 36, wherein the nucleotide sequence is shown in SEQ ID NO. 1-36.
6. Use of the trnG-trnS barcode database of Lycium barbarum samples of claim 5 for identifying varieties of Lycium barbarum.
7. Use of the trnG-trnS barcode database of Lycium barbarum samples of claim 6 for identifying varieties of Lycium barbarum, wherein: the method comprises the following steps: and comparing the trnG-trnS bar code sequence of the sample to be identified with the sequences in the trnG-trnS bar code database to identify the variety of the medlar and determine the interspecific relationship of the medlar.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US17/137,191 US20210198756A1 (en) | 2020-04-28 | 2020-12-29 | Dna barcode for variety identification of wolfberry and identification method therefor |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2020103478862 | 2020-04-28 | ||
| CN202010347886.2A CN111394501A (en) | 2020-04-28 | 2020-04-28 | Method for rapidly identifying medlar based on DNA bar code |
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| CN111690763A true CN111690763A (en) | 2020-09-22 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010347886.2A Withdrawn CN111394501A (en) | 2020-04-28 | 2020-04-28 | Method for rapidly identifying medlar based on DNA bar code |
| CN202010572251.2A Pending CN111893202A (en) | 2020-04-28 | 2020-06-22 | Method for rapidly identifying medlar based on DNA bar code |
| CN202010608982.8A Pending CN111690763A (en) | 2020-04-28 | 2020-06-30 | DNA bar code for identifying variety of Chinese wolfberry and identification method thereof |
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| CN202010347886.2A Withdrawn CN111394501A (en) | 2020-04-28 | 2020-04-28 | Method for rapidly identifying medlar based on DNA bar code |
| CN202010572251.2A Pending CN111893202A (en) | 2020-04-28 | 2020-06-22 | Method for rapidly identifying medlar based on DNA bar code |
Country Status (3)
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|---|---|
| US (2) | US20210198756A1 (en) |
| CN (3) | CN111394501A (en) |
| WO (1) | WO2021218206A1 (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110229927A (en) * | 2019-06-20 | 2019-09-13 | 上海诺德生物实业有限公司 | A kind of method and its application based on DNA bar code identification black fruit fructus lycii |
| CN111394501A (en) * | 2020-04-28 | 2020-07-10 | 宁夏农林科学院枸杞工程技术研究所 | Method for rapidly identifying medlar based on DNA bar code |
| CN113403415B (en) * | 2021-05-31 | 2023-05-26 | 宁夏农林科学院枸杞科学研究所 | Rapid typing identification method of wolfberry S gene based on targeted sequencing |
| CN114292940B (en) * | 2021-12-15 | 2022-11-08 | 西北大学 | Combined barcode sequences and methods for identification of rehmannia and rehmannia plants |
| CN114214336B (en) * | 2022-02-23 | 2022-05-03 | 中国科学院华南植物园 | Lycium ruthenicum LrNOR gene and application of protein thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110229927A (en) * | 2019-06-20 | 2019-09-13 | 上海诺德生物实业有限公司 | A kind of method and its application based on DNA bar code identification black fruit fructus lycii |
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| CN107267640B (en) * | 2017-07-27 | 2020-12-04 | 中国农业科学院特产研究所 | Identification method of vitis amurensis germplasm resources and application thereof |
| CN107523639A (en) * | 2017-09-30 | 2017-12-29 | 数字本草中医药检测有限公司 | A kind of lycium barbarum uses SNP specific primers and its discrimination method with matrimony vine identification |
| CN109486984A (en) * | 2018-08-03 | 2019-03-19 | 广东天保参茸有限公司 | A kind of combination bar code sequence and its accurate identification method identified for ginseng and American Ginseng |
| CN109439787A (en) * | 2018-11-14 | 2019-03-08 | 黑龙江省农业科学院草业研究所 | A kind of molecule labelling method identifying graminous pasture |
| CN110592265B (en) * | 2019-10-29 | 2023-11-24 | 江西省农业科学院蔬菜花卉研究所 | DNA bar code and method for rapid identification of solanum plants |
| CN111394501A (en) * | 2020-04-28 | 2020-07-10 | 宁夏农林科学院枸杞工程技术研究所 | Method for rapidly identifying medlar based on DNA bar code |
-
2020
- 2020-04-28 CN CN202010347886.2A patent/CN111394501A/en not_active Withdrawn
- 2020-06-22 CN CN202010572251.2A patent/CN111893202A/en active Pending
- 2020-06-30 CN CN202010608982.8A patent/CN111690763A/en active Pending
- 2020-12-25 WO PCT/CN2020/139586 patent/WO2021218206A1/en active Application Filing
- 2020-12-29 US US17/137,191 patent/US20210198756A1/en active Pending
- 2020-12-29 US US17/136,208 patent/US20210207226A1/en active Pending
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|---|---|---|---|---|
| CN110229927A (en) * | 2019-06-20 | 2019-09-13 | 上海诺德生物实业有限公司 | A kind of method and its application based on DNA bar code identification black fruit fructus lycii |
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| 石志刚等: "枸杞属植物DNA 条形码研制及其物种的鉴定研究" * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111394501A (en) | 2020-07-10 |
| WO2021218206A1 (en) | 2021-11-04 |
| CN111893202A (en) | 2020-11-06 |
| US20210207226A1 (en) | 2021-07-08 |
| US20210198756A1 (en) | 2021-07-01 |
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